Department of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, China Medical University, No. 91 Hsueh-Shih Rd., Taichung 40402, Taiwan.
Institute of Translation Medicine and New Drug Development, China Medical University, No. 91 Hsueh-Shih Rd., Taichung 40402, Taiwan.
Bioorg Chem. 2024 Jul;148:107424. doi: 10.1016/j.bioorg.2024.107424. Epub 2024 May 3.
Pyrazolopyrimidine derivatives, including pyrazolopyrimidines, 6-aminopyrazolopyrimidines, 6-[(formyloxy)methyl]pyrazolopyrimidines, 6-(hydroxymethyl)pyrazolopyrimidine, and 6-(aminomethyl)pyrazolopyrimidines have been successfully prepared and tested against NCI-H226, NPC-TW01, and Jurkat cancer cell lines. Among the tested pyrazolopyrimidine compounds, we found 6-aminopyrazolopyrimidines and 6-(aminomethyl)pyrazolopyrimidines with essential o-ClPh or p-ClPh substituted moieties on N-1 pyrazole ring exhibited the best IC inhibition activity for Jurkat cells. Furthermore, optimization of the SAR study on the C-6 position of pyrazolopyrimidine ring demonstrated that 6-(N-substituted-methyl)pyrazolopyrimidines 17b, 17d, and 19d possessed the significant IC inhibitory activity for the different leukemia cell lines, especially for Jurkat, K-562, and HL-60. On the other hand, further SAR inhibition and docking model studies revealed that compound 19d, which has a 3-(1H-imidazol-1-yl)propan-1-amino side-chain on the C-6 position, was able to form four hydrogen bonds with residues Ala226, Leu152, and Glu194 and specifically extended into the P1 pocket subsite with Aurora A, resulting in improved inhibitory activity almost similar to SNS-314. To explore the anti-cancer mechanism, compound 19d was measured by Western blot analysis in Jurkat T-cells, however, it showed non-responsibility to Aurora B. For the further structural modifications on the lateral chain of compound 19d, compounds 24 with longer lateral chain were designed and synthesized for testing leukemia cell lines. However, compounds 24 was significantly decrease inhibition potency against leukemia cell lines. Based on the in-vitro results, compounds 17b and 19d could be considered to be the best potential lead drug in our study for the development of new and effective therapies for leukemia treatment. On the other hand, the DHFR inhibition results indicated compound 19d possessed good inhibitory activity and better than the reported naphthalene derivative. Through further comparisons of the model superposition of three-dimensional (3D) conformations in DHFR, compound 19d presented a similar structural alignment to Methotrexate and the reported naphthalene derivative and led to similar drug-like functional relationships. As a results, compound 19d would be a potential DHFR inhibitor for anti-leukemia drug candidate.
吡唑并嘧啶衍生物，包括吡唑并嘧啶、6-氨基吡唑并嘧啶、6-[(甲氧基)甲基]吡唑并嘧啶、6-羟甲基吡唑并嘧啶和 6-(氨甲基)吡唑并嘧啶，已成功制备并针对 NCI-H226、NPC-TW01 和 Jurkat 癌细胞系进行了测试。在所测试的吡唑并嘧啶化合物中，我们发现 N-1 吡唑环上带有必需的邻-ClPh 或对-ClPh 取代基的 6-氨基吡唑并嘧啶和 6-(氨甲基)吡唑并嘧啶对 Jurkat 细胞表现出最佳的 IC 抑制活性。此外，对吡唑并嘧啶环 C-6 位的 SAR 研究进行了优化，结果表明，6-(N-取代甲基)吡唑并嘧啶 17b、17d 和 19d 对不同的白血病细胞系具有显著的 IC 抑制活性，特别是对 Jurkat、K-562 和 HL-60。另一方面，进一步的 SAR 抑制和对接模型研究表明，化合物 19d 在 C-6 位上具有 3-(1H-咪唑-1-基)丙-1-氨基侧链，能够与残基 Ala226、Leu152 和 Glu194 形成四个氢键，并且特别扩展到 Aurora A 的 P1 口袋亚基，从而导致抑制活性几乎与 SNS-314 相似。为了探索抗癌机制，在 Jurkat T 细胞中通过 Western blot 分析测量了化合物 19d，然而，它对 Aurora B 表现出无反应性。为了进一步对化合物 19d 的侧链进行结构修饰，设计并合成了具有更长侧链的化合物 24 以测试白血病细胞系。然而，化合物 24 对白血病细胞系的抑制活性显著降低。基于体外结果，化合物 17b 和 19d 可被认为是我们研究中用于开发新的有效白血病治疗方法的最佳潜在先导药物。另一方面，DHFR 抑制结果表明，化合物 19d 具有良好的抑制活性，优于报道的萘衍生物。通过进一步比较 DHFR 中三维 (3D) 构象的模型叠加，化合物 19d 呈现出与 Methotrexate 和报道的萘衍生物相似的结构排列，并导致类似的类药性功能关系。因此，化合物 19d 可能是一种有潜力的 DHFR 抑制剂，可作为抗白血病候选药物。
